Dataset Information

Arabidopsis thaliana acclimation to fluctuating light

ABSTRACT: Plants are subjected to perpetual fluctuations of light intensity and spectral composition in their natural growth environment, particularly due to movement of clouds and upper canopy leaves. Sudden exposure to intense light is accompanied by absorption of excess light energy, which results in an overload of photosynthetic electron transport chain and generation of reactive oxygen species in and around thylakoid membranes. To cope with this photooxidative stress and to prevent chronic photoinhibition under dynamically changing light intensities, plants have evolved various short- and long-term photoprotective mechanisms. We used quantitative mass spectrometry to investigate long-term acclimation of Arabidopsis thaliana leaf proteome to fluctuating light (FL) which induces photooxidative stress. After three days of FL exposure the proteomes of young and mature leaves were analyzed separately in the morning and at the end of day to examine possible interaction between FL acclimation and leaf development or time of day.

INSTRUMENT(S): maXis

ORGANISM(S): Arabidopsis Thaliana (mouse-ear Cress)

TISSUE(S): Leaf

SUBMITTER: Pitter Huesgen

LAB HEAD: Pitter Huesgen

PROVIDER: PXD015330 | Pride | 2020-01-30

REPOSITORIES: Pride

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Dataset's files

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			Action	DRS
	MaxQuanttxtfolder.zip	Other
	RAWdata.zip	Other

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Publications

Photoprotective Acclimation of the <i>Arabidopsis thaliana</i> Leaf Proteome to Fluctuating Light.

Niedermaier Stefan S Schneider Trang T Bahl Marc-Oliver MO Matsubara Shizue S Huesgen Pitter F PF

Frontiers in genetics 20200305

Plants are subjected to strong fluctuations in light intensity in their natural growth environment, caused both by unpredictable changes due to weather conditions and movement of clouds and upper canopy leaves and predictable changes during day-night cycle. The mechanisms of long-term acclimation to fluctuating light (FL) are still not well understood. Here, we used quantitative mass spectrometry to investigate long-term acclimation of low light-grown <i>Arabidopsis thaliana</i> to a FL conditio ...[more]

PMID: 32194630

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Project description:As part of our study on human MASTL (microtubule-associated serine/threonine kinase-like), we used mass spectrometry (MS)-based proteomics in the aim to unravel novel aspects about MASTL activation, regulation and potential substrates. Specifically, we performed a phosphoproteomics-based kinase assay (essentially as in Xue et al. 2014) to identify MASTL substrates and analyzed different MASTL constructs to generate a “phosphorylation map” of the MASTL protein. siKALIP (Stable Isotope Labelled Kinase Assay-Linked Phosphoproteomics) Three MASTL variants, which we termed MASTL (full-length), MASTL450 (short) and Bonsai (truncated), were expressed in HEK293 cells and isolated using a tandem Strep and His-tag affinity purifications. For the in vitro kinase assay we used a dephosphorylated cell extract from interphase HEK293 cells to perform a slightly modified version of the siKALIP (described in Xue et al. 2014). Samples included an untreated basal sample, a dephosphorylated sample and samples for each of the three MASTL constructs. For label-free quantification, reactions were performed in quadruplicates for each MASTL construct. Explanation for MS raw files with MASTL construct included: FL: MASTL full-length. 450short: Shorter (consisting of the first 450 amino acid residues) MASTL. Bonsai: Truncated MASTL (consisting of amino acid residues 35-112, 113-180 and 728-879). MASTL phosphorylation map To analyse the phosphorylation state of MASTL we used isolated protein from an interphase HEK293 cell culture, either alone or incubated with mitotic extracts from HEK293 cells. Samples of different MASTL constructs were used for in-gel digestion and analysed by MS. Explanation for MS raw files with MASTL construct included: FL: MASTL full-length (“INACTIVE” indicates kinase-dead, “Dephos” indicates dephosphorylated FL protein, “Mit” indicates incubated with mitotic extracts). 450: Shorter (450 amino acids) MASTL (“KD” indicates kinase-dead). BONSAI: Truncated MASTL (“KD” indicates kinase-dead).